Bis quaternary oximes



Unite BIS QUATERNARY UXIMES Brennie E. Haclrley, In, 146B Hawthorne Drive, and Edward J. Pozionlclr, 149E Hawthorne Drive, both of Edgewood, Md., and George M. Steinberg, 3111 Hatton Road, Baltimore 8, Md. No Drawing. Filed Apr. 28, 1959, Ser. No. 869,578 8 Claims. (Cl. 26i)296) (Granted under Title 35, US. Code (1952), sec. 266) OH=NOH CH=NOH (I) where R is a polymethylene group having from two to six carbon atoms and X- is chloride, bromide or iodide.

This invention further includes compounds of Formula I, but in which R is a polymethylene group containing from seven to ten carbon atoms.

This invention further includes 'l, l'-(2,-5-rlimethyl-pphenylenedimethylene) bis (4-formylpyridinium) halide dioximes of the formula OH=NO H CH=NOH (II) The invention also relates to 1,1-(2-butenylene) bis (4-formylpyridinium) halide dioximes of the formula on=NoH CH=NOH (In) The invention further relates to 1,1-polyrnethylene his (3-formylpyridinium) halide dioxirnes of the formula ll on=Non C H=N O H (IVa) wherein R is a polymethylene group containing from 2 to 6 carbon atoms.

Another class of compounds included are the 1,1-(pphenylenedimethylene) bis (3-formylpyridinium) halide dioximes of the formula CH=NOH CH=NOH States Patent This invention further relates to unsymmetrical bis. quaternary 4-formylpyridinium halide monoximes of the formula CH=NOH (V) NH: NHQ This invention also relates to the compounds (VII) The invention also relates to the compounds N:(oHt 3 N U UCH- NOH (VIII) While all these compounds are useful in varying degree for the purposes set out above, the difierent groups exhibit quite striking difierences in effectiveness. In all the above formulas X" is chloride, bromide or iodide, which appear to be equivalents as to physiological action, except for their afiect on solubility.

Wilson et al., in Patent No. 2,816,113, granted Dec. 10, 1957, disclose a group of compounds which are effective as antidotes for treatment of mammals poisoned by compounds of high anticholinesterase activity, such as the nerve gases diisopropylphosphorofluoridate (DFP), isopropyl methylphosphonofiuoridate (GB), and O-ethyl, N,N-dimethyl-phosphoroamidocyanidate (GA), as well as other related organic phosphorus compounds, including many insecticides. The compound of the Wilson et a1. group which has received by far the most attention is 2-formyl1-methy1 pyridinium iodide oxime, commonly known as Z-pyridine aldoximemethiodide or Z-PAM. This compound is outstanding in its ability to reactivate, in vitro, acetyl cholinesterase which has been inhibited by, for example, GB. Thus, even as compared to the very closely related 4-PAM, disclosed in Example H of the Wilson patent, 2-PAM shows much greater activity. With isopropyl methylphosphonylated acetylcholinesterase the rate constant at pH 7.4 and 25 C. in the presence of 10* M acetylcholine is 2x10 per mole per minute for Z-PAM and 1.4x10 per mole per minute for 4-PAM. The Wilson patent shows a high rate of survival in mice which had been poisoned with paraoxon and then treated with Z-PAM.

Nerve gas poisoning has been treated symptomatically with drugs Which are pharmacologically antagonistic to acetyl choline. Such a compound is atropine audit is at present the recommended remedy. Recently, 2-PAM has been reported to enhance considerably the activity of atropine in the chemotherapeusis of poisoning due to organophosphorus compounds.

The compounds of Formula I above in which R contains from 2 to 6 carbon atoms are appreciably more TABLE 1 R 3 Rate constant (llmoles/minutes) (CH2): 7X10 (CH2): 6X10 M 6X10 1x10 (CH2); (5X10 When administered in combination with atropine to animals poisoned with GB the order of eitectiveness was somewhat different. Under these conditions the compound in which R=(CH i.e., 1,1'-trimethylene bis (4-formylpyridinium) bromide dioxime also known as .TMB-4, was most effective. In rats challenged with a 2LD dose of GB administered intravenously, all of a group of six animals survived if the atropine-TMB-4 combination was administered intravenously immediately after poisoning. The atropine-2-PAM combination saved only two of the group of animals. On the other hand, with dogs which were given a 20LD dose of GB subcutaneously the survival ratios were the same (4/5) for the two treatments, which were given intravenously when symptoms appeared. However, the recovery time was much shorter for the surviving animals which received the TMB-4, i.e., 2 hours, as against 24 hours for those receiving the 2-PAM.

. A summary of the reactivation rates and survival ratios for these compounds when administered therapeutically to rats together with atropine is as follows:

TABLE 2 R Reactivation Survival rate constant ratio (GB) (CH2): 7X10 6/6 (012193 6X10 6/6 (SE9 6X10 (CHM 1x10 6/6 0112 6x10 3/4 Mice 0.173 rug/kg. (LDgg). Rats 0.126 mg./kg. (2LD Rabbits:

Intravenous 0.340 mg./kg. (ZOLD Subcutaneous 0.900 mg./kg. (20LD Dogs and cats:

Intravenous 0.440 mg./kg. (20LD Subcutaneous 0.900 mg./kg. (20LD To minimize absorption effects both the GB and TMB-4 were ordinarily given intravenously. However, in the therapeutic tests on rabbits, dogs and cats, the GB was administered subcutaneously, since death from 20LD intravenous dose of GB occurs so quickly that it is virtually impossible to give timely administration of the antidote.

Atropine, when administered, was included in the following amounts.

Rats 4 mg./kg. Rabbits 2 mg./kg. 5 Dogs and cats 0.5 rug/kg.

.The prophylactic doses were given withintwo minutes prior to the injection of the GB, the therapeutic doses so soon as poisoning symptoms were visible. 10 Table 3 shows the results.

TABLE 3 A. PROPHYLATIC 2-PAM TMB-4 Animals Survial ratio Survival ratio Dose,

Dose, mg./kg.

Without With atropine atropine Without With atropine atropine Mice.-. 40 12 0/10 Rats. 40 1/6 1/6 6/6 Rabbits. 5 2/6 5 4/6 Cats 40 5/5 20 8/5 Dogs 40 4/5 20 4/5 The recovery periods, i.e., time for disappearance of symptoms of poisoning, among survivors in the above tests, with atropine, were as follows.

TABLE 4 2-IAM TMB-4 Animals Prophy- Thera- Prophy- Theralactic peutic lactic peutic ats 60 min 15 min. Rabbits 3 hr 30 min 2 hr. r Cats. 5 hr. 5 hr. 5 hr 24 hr. 40 Dogs 24 hr 24 hr 1% hr- 3 hr.

The compounds of Formula I in which R contains from 7 to 10 carbon atoms are less effective than those of our preferred group. For these compounds the reactivation rate constant and the survival ratio for rats (measured as given above) were as follows, X'" being bromide.

TABLE 5 Survival R Rate constant ratio zh 2x10 0/4 (ones 1 2x10 0 4. 011i) 0/4 (0112M 1 4x10 016 While these compounds were ineltective in vivo against GB, they were, together with Z-PAM, very effective against certain other anticholinesterases, particularly that designated as VX by the U.S. Army Chemical Corps. All these compounds caused survival of all animals (survival rates of 4/4 and 6/6), when administered therapeutically to rats challenged by 2LD doses of VX.

The compounds of Formula II exhibited properties in termediate those of the two subgroups of Formula I. When X- was chloride the compound had the following properties. (In this and all following tables the survival ratios are those for rats challenged by 2LD doses of GB or (VX) and the oxime was employed therapeutically.)

Compounds of Formula III showed reactivation rates very close to those of ourpreferred group. Thus when X in Formula 111 is bromide the reactivation rate constant was 8x10 as compared to the value for the R=(CH member of our preferred group of 6x10 For the unsaturated member (Formula III) the survival ratio for rats challenged by GB was only 1/4 as compared to 6/6 for the saturated analogue (Formula I). Both gave complete survival (ratios of 4/4 and 6/6) for animals challenged by VX, however.

Compounds of Formula IVa showed anomalous properties.

They gave reactivation rates which were low, but survival ratios which were high as compared to Z-PAM, as shown by Table 6, X being bromide.

TABLE 6 Reactivation Survival R rate constant ratio The compounds of Group IVb, which are closely related to those of IVa, were somewhat less effective. When X- was bromide the compound had the following properties: Reactivation Rate Constant 2x10 Survival Ratio (GB)--2/4.

The compounds of group V were another group in which the results of therapeutic treatment against GB were better as compared to 2-PAM then the reactivation rate constants would suggest, as shown by the following table, X- being bromide.

TABLE 7 Reactivation Survival REN rate constant ratio The compounds of Formulas VI, VII and VIII, while being of dilferent structure are alike in exhibiting reactivation rate constants which are very low as compared to 2-PAM but giving high survival ratios as shown by Table 8, X- being bromide in each case.

TABLE 8 Formula No. Reactivation Survival r e ratio (GB) 69 4/4 Negligible 4/4 67 4/4 Preparation of Compounds priate omega-halopropyl quaternary salt in a 1.5:1 molar ratio. Two procedures were utilized.

Procedure A.A mixture of the pyridine oxime and halide was dissolved in sufiicient ethanol and refluxed for the period of time specified in Table 9.

Procedure B.A mixture of the oxime and halide was dissolved in about 100 ml. of ethanol and heated in a 200 ml. capped pressure bottle (carbonated beverage type) for the length of time specified. The reaction mixtures were cooled to room temperature and the product removed by filtration. In several instances it was necessary to add absolute ether to effect complete precipitation. The products were recrystallized from ether. This procedure was usually employed because of its simplicity.

Table 9 gives the procedure, yields and melting points for representative compounds.

TABLE 9 SUBSTITUENTS Melting Formula Coudi- Yield, (m) or de- No. tions percent eomposi Ha- R R" tion ((1) lide point, 0

(CH2); A, 31 hr 35.0 300 111 (CI-Ir): B, 48 hr-- 88. 2 238-241 6. (CH2); B, 16 hr- 81.0 239-241 (1 (CEIm B, 95 hr.. 95. 0 208-210 d I (CH2)10 B, 8 r 85.0 219-223 (1 II B, 68 hr.. 70 300 111 (GHm B, 60 hr.- 68 208-211 111 IV/z Br (0512):: B, 60hr 226-231 ml IVb Br B, 20 hr 83. 5 248-251 m V Br (C H5)3 B, 69 hr.- 43 230-231 d. V Br Pyridine B, 64 hr.- 223-226 d.

ring. VIII Br B, hr-. 16 201-203 (1.

Further details regarding the preparation and properties of certain of our compounds are given in the following publications, by us and our associates:

Pyridine Aldoximes by Edward I. Poziomek, Brennie E. Hackley, Jr. and George M. Steinberg, Journal of Organic Chemistry," vol. 23, pp. 714-717, (May 1958); and Chemotherapeutic Effectiveness of Trimethylene Bis (4-Formy1 Pyridinium Bromide) Dioxime in Anticholinesterase Poisoning by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society for Experimental Biology and Medicine, vol. 98, pages 107-109 (May 1958). These articles are to be considered incorporated by reference in this specification.

While we have shown a number of specific examples of compounds and their use, it will be obvious that various changes can be made without departing from our invention, which is defined by the following claims.

We claim:

1. A 1,1'-polymethylene bis(4-formyl pyridiniurn) oxirne halide compound of the formula T N [D R (3X -7 s 8. A 1,1 polymethylene bis-(3-formylpyridinium) References Cited in the file of this patent halide dioxirne Of the formula UNITED STATES PATENTS R 2,816,113 Wilson et a1 Dec. 10, 1957 5 2,819,997 McLamore Jan. 14, 1958 2,909,527 Shapiro et a1. Oct. 2 0, 1959 OH=NOH OH=NOH 2,924,604 Steinhards et a1. Feb. 9, 1960 wherein R is a polymethyiene containing from 2 to 6 FOREIGN PATENTS carbon atoms and X- is selected from the class consisting Poziornek et a1.: Abst. of Papers, 132ml Meeting of -of chloride, bromide and iodide. 10 ACS, August 27, 1957. 

1. A 1,1''-POLYMETHYLENE BIS(4-FORMYL PYRIDINIUM) OXIME HALIDE COMPOUND OF THE FORMULA
 8. A 1,1'' - POLYMETHYLENE BIS-(3-FORMYLPYRIDINIUM) HALIDE DIOXIME OF THE FORMULA 